Assembly PCR Primer Designer (Beta)

Design assembly PCR primers for NEBuilder HiFi, GeneArt, In-Fusion and overlap PCR.

Assembly primer inputs

Assembly setup

Includes 1 vector + 2 inserts.

15–20 bp guidance for 3 total fragments; editable.

Vector

PCR opens after this base. Leave blank for an already-linear, blunt-ended vector.

Insert fragments

Enter fragments in assembly order, without added overlap tails.

Primer settings

Reaction settings

References

Kit guidance and primer Tm sources.

  1. Assembly guidance: NEBuilder HiFi, GeneArt Seamless and In-Fusion Snap manuals.
  2. Primer Tm: SantaLucia (1998); salt correction: Owczarzy et al. (2008).